Level Of Growth Inhibition Research Articles

Synaptotagmin-1 antagonizes paraquat intracellular accumulation and nephrocyte toxicity by up-regulating SERBP1/GLUT2 expression

Acute kidney injury (AKI) is common and an independent risk factor for mortality in patients with paraquat (PQ) poisoning. Currently, no specific antidote is available. Synaptotagmin-1 (SYT1) has been identified as a key protein that facilitates PQ efflux in PQ-resistant A549 cells, thereby preventing PQ-induced lung injury. However, the protective effect of STY1 on PQ-induced AKI remains to be elucidated. This study exposed human kidney 2 (HK-2) cells overexpressing SYT1 to PQ. These cells exhibited significantly lower levels of growth inhibition, reactive oxygen species production, early apoptosis, and PQ accumulation compared to the parent HK-2 cells. Transcriptomic screening and Western blot analysis revealed that SYT1 overexpression significantly promoted the expression of glucose transporter 2 (GLUT2). Inhibition of GLUT2 completely abolished the protective effects of SYT1 overexpression in HK-2 cells and restored intracellular PQ concentrations. Further immunoprecipitation-shotgun and RNA interference experiments revealed that SYT1 binds to and stabilizes the protein SERPINE1 mRNA-binding protein 1 (SERBP1), enhancing the stability of GLUT2 mRNA and its protein levels. In summary, SYT1 antagonizes PQ intracellular accumulation and prevents nephrocyte toxicity by up-regulating SERBP1/GLUT2 expression. This study identifies a potential target for the treatment of PQ-induced AKI.

EVALUACIÓN DE ESTRATEGIAS DE MANEJO ECOLÓGICO DE ANTRACNOSIS DE LA PAPAYA EN CAMPO Y POSCOSECHA

<p><strong>Background:</strong> Mexico is the third producer and the first exporter of papaya in the world. The crop has multiple phytosanitary limitations, among which anthracnose stands out, which can cause substantial production losses. The main management method is with chemical synthesis fungicides. <strong>Objective:</strong> To evaluate treatments with biorational characteristics to control anthracnose in papaya crops in pre- and post-harvest. <strong>Methodology:</strong> The causal agent of anthracnose was isolated and identified, the growth inhibitory effect of<em> </em><em>C. gloeosporioides</em><em> </em>was evaluated <em>in vitro</em> with extracts of <em>Datura stramonium</em>, <em>Argemone mexicana</em>, <em>Citrus sinensis</em> essential oil and <em>Eucalyptus globulus</em> essential oil; In addition, biorational treatments were applied pre- and post-harvest (botanical extracts, essential oils and <em>Trichoderma harzianum</em>); The incidence and severity were evaluated and the percentage of efficacy was calculated. A randomized complete block design was used and analysis of variances and comparisons of means were performed (Tukey, P ≤ 0.05). <strong>Results:</strong> The <em>E. globulus</em> oil extract reached the highest in vitro growth inhibition levels, followed by the <em>C. sinensis</em> essential oil and the <em>A.</em><em> mexicana</em> extract; In the field phase, <em>T. harzianum</em> reached 80.7% efficacy and 8% incidence, the essential oil of <em>E. globulus</em> and the essential oil of <em>C. sinensis</em> reached 73 and 66% efficacy, respectively. The effect of the treatments applied in pre-harvest demonstrated that successive applications of <em>T. harzianum</em> reduced severity by 81% and incidence on papaya fruits by 30%, the essential oil of <em>E. globulus</em> and that of <em>C. sinensis</em> achieved efficacy of 64 and 68%, in their order. Finally, in the storage phase, <em>T. harzianum</em> achieved a control efficiency of 89%, <em>C. sinensis</em> oil 84% and 85% for the essential oil of <em>E. globulus</em>, <em>A. mexicana</em> managed to inhibit 76.46%. <strong>Implications:</strong> Comparison of other biorational management treatments such as the use of essential elements (silicon, sulfur, etc.), nanotechnology, UV irradiation and resistance inducers is suggested. <strong>Conclusion:</strong> These results support that a successful and biorational control of anthracnose in papaya must be based on comprehensive management that considered the preharvest and storage phases.</p>

Predicting epirubicin response in Danish patients with breast cancer.

539 Background: Anthracyclines are highly effective chemotherapy drugs. However, they are toxic and feared for their cardiotoxicity. There is a need for improved personalized prediction of treatment benefit. For example, state-of-the-art guidelines (NCCN Guidelines Version 1.2024 and meta-analysis PMID: 37061269) mark anthracyclines as optional adjuvant treatment of estrogen receptor (ER) positive, HER2 negative early high-risk breast cancer. A biomarker to predict whether a patient will benefit from anthracycline treatment could potentially lead to increased cure rate and overall reduced toxicity. However, no such test exists. Here we introduce a clinically validated, predictive test based on clinical tumor transcriptome data and the cell line panel NCI60 that can forecast epirubicin response in breast cancer patients with high accuracy. Methods: Microarray gene expression levels and growth inhibition levels (GI50) from the NCI60 cell line panel were acquired from NCBI to build a prediction algorithm. A tumor-specific variance-based filter based on >6.000 patient tumors was applied to select the most relevant genes. AUC (Area Under Curve) was used to evaluate the performance of numerous configurations and combinations of parameters that led to different gene-signature sets. Response scores based on the gene expression level of all genes in each signature were generated for each sample in a published cohort (PMID: 23340299) and a final signature with 141 genes was the best-performing signature (Progressive Disease versus Response AUC 0.81 and ANOVA-test p-value < 0.01). The model was validated on a cohort of Danish breast cancer patients treated with epirubicin in the metastatic setting (n=153), with time-to-progression (TTP) as endpoint (PMID: 31654283, PMID: 30099635). Finally, we used an established method (PMID: 30289602) to test if the gene signature was correlated between microarray- and RNA sequencing-based datasets. Results: The hazard ratio (HR) for the best-performing signature was 0.31 when comparing patients with a difference of at least 50 in response score (two-sided p-value = 0.012, 95% confidence interval 0.12-0.77, endpoint TTP, continuous scoring). This was consistent in a multivariate model including ER and HER2. Using the data and method provided by Pedersen et al (PMID: 30289602), we also saw a high correlation between matching microarray and RNA sequencing datasets for the genes in our model (mean Pearson’s r = 0.87). Conclusions: Here is presented a development and clinical validation of an accurate epirubicin response signature for breast cancer patients. The expression of the genes in the signature is highly correlated between microarray- and RNA sequencing data. Moving forward, we will seek to validate the model independently on RNA sequencing data. Our aim is that these models will be accepted for prospective clinical tests at local hospitals in Denmark in early 2025.

RHIZOBACTERIA AS ANTAGONIST AGAINST Fusarium oxysporum CAUSING TOMATO WILT

Tomatoes (Lycopersicon esculentum L.) are the most extensively grown vegetable in the world. It belongs to the Solanaceae family and is commonly planted for its tasty fruits. Pests, weeds, diseases, and parasites are just a few of the numerous variables that significantly affect tomato growth and yield. The most common disease affecting tomatoes is fusarium wilt. Fifteen rhizobacterial strains were identified by morphological and biochemical analyses in this work, and they were employed as an antagonist against Fusarium oxysporum f.sp. lycopersici. Fifteen isolates were investigated for their antagonistic properties against Fusarium oxysporum f.sp. lycopersici using an invitro dual culture approach. The growth of Fusarium oxysporum f.sp. lycopersici was suppressed by each isolate. Out of the 15 rhizobacteria isolates, isolate RBS-5 exhibited the highest level of growth inhibition and strongly suppressed the development of Fusarium oxysporum f.sp. lycopersici, resulting in a 57.28 percent reduction in pathogen growth as compared to the control. The development of Fusarium oxysporum f.sp. lycopersici was suppressed by isolates RBS-12, RBS-6, and RBS-15, in decreasing order of merit, compared to the control by 53.7, 51.91, and 51.73 percent. Isolate RBS-13 showed the least amount of pathogen growth inhibition 20.83 percent. The data was statistically analyzed.

Phytotoxicological assessment of AMD affected river waters in “Sredna Gora”, Bulgaria to Lepidium sativum L.

The ongoing pollution of water bodies from decommissioned mines and mining waste facilities in Bulgaria is a serious problem, leading to a contamination of river waters with heavy metals. Although the chemical analyses of acid mine drainage (AMD) affected waters provide information about their toxicity, this information is not enough to evaluate the potential impacts on the biota. For this, different biological responses are used to assess the toxicity of the polluted waters.The goal of the research was to determine the phytotoxicity (using Lepidium sativum) of five affected river waters. The lower reaches of the rivers flow though agricultural areas and are often used for irrigation by local farmers, increasing the chance of produce contamination.The effect of the working solutions on seed germination and primary root growth of L. sativum was assessed by the following indices: Relative seed germination (RSG), relative root growth (RRG) and germination index (GI, the product of RSG and RRG).The obtained results have shown the connection between germination/root growth inhibition and concentrations of heavy metals in working solutions, but there is no connection between germination/root growth inhibition and pH levels.

Alternaria brassicicola’ya karşı Trans-cinnamic Asit ve Xenorhabdus szentirmaii’nin Antifungal Metabolitlerinin Kullanım Potansiyellerinin Araştırılması

The, antifungal activities of trans-cinamic acid (TCA) and cell-free supernatant (CFS) of X. szentirmaii were evaluated against Alternaria brassicicola that cause Black spot disease. The results showed that TCA was more suppressive than CFS of X. szentirmaii in the control of A. brassicicola. In petri assays, the highest suppressive effect on spore germination was in TCA treatments compared to CFS of X. szentirmaii. Moreover, no germ tube elongation was observed in any of tested concentrations (0.25%, 0.50%, 1%, 2%) of TCA. The highest level of mycelial growth inhibitions (100% and 92%) were exhibited by TCA (2%) and TCA (1%), respectively. The curative and protective activity of TCA on disease severity of A. brassicicola were also evaluated on broccoli plants. In the curative activity assays, disease severities were 71.00%, 56.33% and 54.03% for control, TCA (1%) and TCA (2%), respectively. Whereas in the protective activity, TCA (2%) suppressed the disease severity significantly greater than TCA (1%). The disease severity of A. brassicicola after TCA application on broccoli plants was 25.21% and 17.37% for TCA (1%) and TCA (2%) respectively. Similar data were obtained in the efficacy of TCA on A. brassicicola. In the curative activity assays, TCA (1%) and TCA (2%) exhibited the efficacy with rates of 20.57% and 23.02%, respectively. However, TCA (2%) showed significantly higher efficacy than TCA (1%) in the protective activity. The current study provides that TCA has a great potential to suppress A. brassicicola and can be a good alternative to synthetic fungicides.

STUDIES ON NOVEL METAL COMPLEXES OF COUMARIN SCHIFF BASE: SYNTHESIS, STABILITY CONSTANT, ANTIMICROBIAL AND ANTICANCER ACTIVITY

A facile synthetic protocol was employed to synthesize new coumarin Schiff base (E)-3-(1-((2-hydroxy-5- methylphenyl)imino)ethyl)-2H-chromen-2-one] and their Co and Cu metal complexes. Synthesized molecules were characterized by various spectroscopic methods. The results of antibacterial studies demonstrated that the maximum level of growth inhibition for S. aureus and B. subtilis was shown by compound 5, with inhibition zones of 16±1 mm and 18.5±1.3 mm, respectively, while, Compound 4 exhibited 12.5±0.5 mm and 14±0.25 mm, respectively. Invitro Cytotoxicity of Cu-complex 5 towards human hepatocellular adenocarcinoma cancer cells, at concentrations of 12.5, 25, 50, and 100 g/mL, displayed a reduction in the percentage of cell viability in a dose-dependent manner, with values of 72.45%, 60.12%, 48.69%, and 23.25%, respectively. The results revealed that Compound 5 has the potential to interact with the bacterial membrane surface, leading to the disruption of the same. Additionally, potentiometric analysis was used to estimate the proton-ligand dissociation constant of HL as well as its metal stability constants with Co2+ and Cu2+ .

NVG-222: A First-in-Class Autoregulating Half-Life Extended ROR1xCD3 T Cell Engager Heralding a New Class of Safer Drugs

Background T cell engagers (TCEs) are realizing their promise in both liquid and solid tumours, with several approvals in the last two years and many more in clinical development. However, the mechanism of action (MOA) that drives efficacy and the impressive clinical response rates comes at a cost. Activation and inflammatory signals induced by TCEs can push T cells into overdrive, resulting in grade 3+ treatment related adverse events (TRAEs). While step up dosing and pre-medication can help in limiting the severity of toxicity, there is a clear clinical need for safer TCEs with a lower frequency of high grade TRAEs to improve tolerability and widen the therapeutic index, which may ultimately result in better efficacy and responses. Our autoregulation (AR) platform technology, applicable across different biologic modalities and multiple indications, enables the deactivation of a drug in response to biological cues emanating from the MOA but only when levels associated with a high rate of toxicity are reached. AR's threshold-based, response-mediated negative feedback loop has the potential to improve safety without affecting potency and is based on similar physiological regulatory pathways that maintain homeostasis. Objective Establish proof-of-concept with AR in the context of NVG-222, our next generation half-life extended (HLE) ROR1-targeting TCE. Methods AR peptides sensitive to the proteolytic activity of immune cell specific protease granzyme B were carefully selected to maintain integrity at the low concentrations of granzyme B associated with TCE potency, whilst undergoing proteolysis at the high concentrations reported in patients with grade 3+ TRAEs. These peptides were embedded in the linker between both targeting arms of ROR1xCD3 TCEs formatted as tandem scFvs. Cytotoxic response and cytokine release potential of TCEs with and without AR were tested in a T cell coculture assay using the ROR1 + Mantle Cell Lymphoma cell line JeKo-1. Safety and efficacy was evaluated in vivo using a model of severe cytokine-induced toxicity in NOD-Scid gamma (NSG) mice engrafted with a ROR1 + solid tumour of MDA-MB-231 cells and reconstituted with human T cells. PK was assessed following a single intravenous injection in Tg32 mice. Results Selected AR peptides were exposed to a range of proteases present in circulation or in the tumour microenvironment and showed a specific and threshold-dependent sensitivity to granzyme B. A first prototype of an AR TCE was engineered from NVG-111, a ROR1xCD3 TCE in clinical development (NCT04763083), and showed uncompromised cytotoxic potency in vitro with an EC 50 in the pM range. This indicates that insertion of a granzyme B sensitive AR peptide does not lead to premature inactivation of the drug. Further evaluation in vivo showed that exposure of tumour-bearing mice to high levels of parental NVG-111 induced severe symptoms of cytokine-induced toxicity, including rapid weight loss, that led to the ethical euthanizing of mice. In contrast, no symptoms of toxicity were observed in the cohort exposed to high levels of AR NVG-111, with all mice surviving to the end of the study. Both parental and AR NVG-111 induced the same level of tumour growth inhibition. The cohort of mice treated with AR NVG-111 had a low or undetectable rate of metastasis. AR technology was then applied to our next generation HLE TCE NVG-222, which builds on the clinical success of NVG-111 by using the same structure at its core, but with a longer half-life that provides more convenient dosing. The cytotoxic potency of AR NVG-222 was shown to be largely equivalent to that of AR NVG-111 (EC 50=38pM vs 13pM respectively), suggesting that similar clinical efficacy should be observed with equivalent dosing. Furthermore, a PK study in Tg32 mice, known to correlate to human PK, showed a t 1/2 of ~12 days and further allometric scaling suggests a potential t 1/2 of ~14 days in clinic. Conclusion Our AR platform technology has the potential to improve the therapeutic index of immunotherapies leading to improved safety and efficacy outcomes. This technology has been applied to our next generation ROR1-targeting TCE NVG-222, which is in IND-enabling studies with the aim of beginning clinical development for hematological and solid malignancies in 2024.

COMPARISON OF ISOLATES OF Phytophthora colocasiae Raciborski FROM DIVERSE ALTITUDES AND APPRAISAL OF PLANT EXTRACTS FOR ITS MANAGEMENT IN VITRO

Leaf blight disease caused by Phytophthora colocasiae Raciborski is a serious constraint to taro production. This study was carried out to characterize the isolates of P. colocasiae from Makurdi, Nigeria and Befang, Cameroon; and evaluate the potential of aqueous leaf extracts of Eucalyptus, neem and mahogany at 0, 50 and 100% concentrations for management of the pathogen. A synthetic fungicide (Mancozeb) was included as a check. The isolate from Befang had more radial growth than that from Makurdi. The colonies of both isolates were translucent and low on corn meal agar (CMA) but opaque and high on acetate differential agar enriched with dextrose (ADAD). The texture of the Befang isolate was filamentous on CMA but hard and grainy on ADAD; the Makurdi isolate was filamentous on both media. Mancozeb and all the plant extracts inhibited the growth of the pathogen significantly (P≤0.05). Among the extracts, Eucalyptus at 100% concentration was the most potent (94.4-100.0% inhibition) and it was comparable to Mancozeb (100% inhibition); mahogany at 50% was the least potent (<20%). Both concentrations of neem gave similar levels of growth inhibition (mean=50.0–64.6%) throughout. Other inhibitory levels were 32.0–57.3% for Eucalyptus at 50% and 24.0–53.9% for mahogany at 100%. These results indicated that ADAD and CMA were appropriate culture media for the fungus. The extracts of Eucalyptus, neem and mahogany had fungicidal activity against P. colocasiae; therefore they should be incorporated into management programmes for the pathogen.

Effect of Different Concentrations of Nano-Fungicides on Mycelial Radial Growth of Phytophthora infestans

Potato (Solanum tuberosum L.) popularly known as the “King of Vegetables” is a starchy, tuberous crop, that belongs to the family Solanaceae. Potatoes play an important role in the daily human diet because they are a copious source of minerals, vitamins, antioxidants and micronutrient. The potato is a basically temperate crop, but it grows under a wide range of climatic conditions. Potato crop is generally affected by different types of biotic and abiotic diseases. Among all diseases, Late blight of potato, caused by the Phytophthora infestans (Mont) de Barry is one of the most devastating diseases of potato worldwide. In India, the losses caused by the disease are 10-20% in Uttar Pradesh, 10-15% in West Bengal and Punjab, 10-15% in Karnataka and Uttrakhand also have been reported during 2013-14. The management of the disease can be done through cultural, mechanical, use of the resistant variety, biological and chemical, etc. The efficacy of current strategies for the management of disease is limited. Considering the destructive nature of pathogen and lack of efficient control measures, the development of alternative or complementary approaches for the management of this disease is highly desirable. However reported use of the same chemicals may raise resistant strain among the pathogens. Therefore, the use of Nano-fungicidal control of spot blotch disease cannot be overstated. However, the disease can be managed with various nanofungicides under in vitro conditions assessed for their efficacy against the disease. The effect of four nanofungicides i.e., Agritec nanofungicide, Silver nanofungicide, Selenium nanofungicide and Nickel nanofungicide against P. infestance was evaluated at different concentrations (50, 75 and 100 ppm) by poison food technique. The in vitro experiment revealed a noticeable reduction in the radial mycelial growth of P. infestance as the concentration of nanofungicides increased. Specifically, at 7 days after inoculation (DAI), the following nanofungicides at a concentration of 100 ppm exhibited the highest levels of mycelial growth inhibition when compared to the control: Silver nanofungicide (72.45%), Agritec nanofungicide (70.21%), Selenium nanofungicide (68.77%), and Nickel nanofungicide (64.65%).

Biological production of epicoccamide-aglycone and its cytotoxicity

Epicoccamide (EPC) is an O-d-mannosylated acyltetramic acid of Epicoccum origin and is a bolaamphiphilic fungal polyketide. EPC displays weak toxicity against Staphylococcus aureus and HeLa cell lines. The EPC biosynthetic gene cluster was previously identified in Epicoccum nigrum and knockout of the glycosyltransferase gene (epcB) abolished EPC production. EPC-aglycone was expected in the epcB knockout but was not found. This study demonstrates that extractive culture using the hydrophobic resin Diaion HP-20 resulted in the production of EPC-aglycone, which was isolated using chromatographic separation techniques, and its structural identity was substantiated by chemical analyses. EPC-aglycone displayed strong antibacterial activity against Staphylococcus aureus, with the minimal inhibitory concentration of 1 μg/mL (64 μg/mL for EPC). EPC-aglycone displayed higher levels of growth inhibition against HeLa cell line (the half inhibitory concentration, 19 μM) and WI-38 (15 μM) cell line than EPC (76 μM and 38 μM vs. HeLa and WI-38, respectively). The dose–response curve fit of growth inhibition indicated that EPC-aglycone adopted a shallow curve (low slope factor), which was different from that of EPC, suggesting that their cellular targets are distinct from each other. This study substantiates that the d-mannose attachment is the final step in EPC biosynthesis, showcasing a glycosylation-mediated modulation of the biological activity of simple acyltetramic acid. This study also highlights the usefulness of extractive cultures in mining cryptic microbial natural products.

Paclitaxel prodrug-encapsulated polypeptide micelles with redox/pH dual responsiveness for cancer chemotherapy

Polypeptides are a highly promising carrier for delivering hydrophobic drugs, due to their excellent biocompatibility, non-toxicity, and non-immunogenicity. Herein, a redox and pH dual-responsive poly(ethylene glycol)-SS-b-polypeptide micelles encapsulated with disulfide bridged paclitaxel-pentadecanoic acid prodrug was developed for cancer chemotherapy. First of all, disulfide bridged paclitaxel-pentadecanoic acid prodrug (PTX-SS-COOH) and poly(ethylene glycol)-SS-b-polylysine-b-polyphenylalanine (mPEG-SS-b-PLys-b-PPhe, ESLP) were synthesized and confirmed via NMR, MS, FT-IR or GPC. After that, PTX-SS-COOH (PSH) embedded mPEG-SS-b-PLys-b-PPhe (ESLP/PSH) micelles were prepared by mixing method based on electrostatic interactions and hydrophobic forces. For comparison, mPEG-b-PLys-b-PPhe (ELP) was mixed with PTX-SS-COOH to generate another kind of micelles (ELP/PSH). The characterization of ESLP/PSH micelles through dynamic light scattering (DLS) and transmission electron microscopy (TEM) revealed a spherical structure with a diameter of approximately 170 nm. It is noteworthy that ESLP/PSH micelles displayed a high drug-loading rate of 22.84%, and excellent stability, which can be attributed to the specific interactions between the prodrug and copolymer. Drug release analysis demonstrated that the micelles exhibited a substantial release of PTX in the presence of GSH at pH 5.0, indicating a pH and redox dual responsiveness. In vivo pharmacokinetic study revealed the ESLP/PSH micelles had increased bioavailability and an extended circulation time. Ultimately, antitumor efficacy and systemic toxicity evaluation in 4 T1 tumor-bearing mice confirmed that ESLP/PSH micelles achieved the highest level of tumor growth inhibition (ca. 83%) and the lowest systemic toxicity in comparison with ELP/PSH micelles and commercialized Taxol®. Taken together, the dual responsive micelles represent a promising PTX formulation with potential clinical application in cancer chemotherapy.

Natural Oils Enhance the Topical Delivery of Ketoconazole by Nanoemulgel for Fungal Infections.

Nanoemulgel (NEG) pharmaceutical formulations are gaining popularity because of their ability to serve both as a nanoemulsion and as a gel. These products are well-known for their ease of use, spreadability, controlled release, and ability to hydrate dry skin. Natural essential oils have been shown to promote the cutaneous permeability of topical formulations, enhancing medication safety and efficacy. Herein, we developed NEG for the enhanced permeation of ketoconazole against candidiasis using clove oil (clove-oil-NEG) or eucalyptus oil (eucalyptus-oil-NEG), using the gelling agents carbopol 943 and hydroxypropyl methylcellulose (HPMC). We tested various excipients to increase the solubility of ketoconazole and formulate a nanoemulsion (NE). We measured the NE droplet particle size, shape, entrapment efficiency, and drug release. Furthermore, the physicochemical properties of the optimized nanoemulsion formulation were characterized by techniques such as Fourier transform infrared (FT-IR) spectroscopy and X-ray diffraction (XRD) analysis. The NEs were loaded into gels to form NEGs. NEGs were characterized for drug content, homogeneity, rheology, spreadability, and antifungal activity against Candida albicans, both in vitro and in vivo. Optimized ketoconazole NEG preparations consisted of either 15% clove oil or 20% eucalyptus oil. Droplet sizes in the optimized NEs were <100 nm, and the polydispersity indexes were 0.24 and 0.26. The percentages of ketoconazole released after 24 h from the clove-oil-NEG and eucalyptus-oil-NEGs were 91 ± 4.5 and 89 ± 7%, respectively. Scanning electron microscopy (SEM) showed that the NEGs had a smooth, uniform, and consistent shape and internal structural organization. The drug contents in the clove-oil-NEG and eucalyptus-oil-NEG were 98.5 ± 2.2 and 98.8 ± 3.4%, respectively. Permeation values of ketoconazole from clove-oil-NEG and eucalyptus-oil-NEG were 117 ± 7 and 108.34 ± 6 μg cm-2, respectively. The ketoconazole NEG formulations also had higher levels of fungal growth inhibition than a marketed formulation. Finally, in vivo studies showed that the NEGs do not irritate the skin. Ketoconazole NEG with either 15% clove oil or 20% eucalyptus oil is stable with better efficacy than ketoconazole alone due to excellent dispersion, drug dissolution, and permeability and thus might be recommended for the effective and safe treatment of candidiasis.

Arabidopsis Topless-related 1 mitigates physiological damage and growth penalties of induced immunity.

Transcriptional corepressors of the Topless (TPL) family regulate plant hormone and immunity signaling. The lack of a genome-wide profile of their chromatin associations limits understanding of the TPL family roles in transcriptional regulation. Chromatin immunoprecipitation with sequencing (ChIP-Seq) was performed on Arabidopsis thaliana lines expressing GFP-tagged Topless-related 1 (TPR1-GFP) with and without constitutive immunity via Enhanced Disease Susceptibility 1 (EDS1). RNA-Seq profiling of the TPR1-GFP lines and pathogen-infected tpl/tpr mutants, combined with measuring immunity, growth, and physiological parameters was employed to investigate TPL/TPR roles in immunity and defense homeostasis. TPR1 was enriched at promoter regions of c. 1400 genes and c. 10% of the detected binding required EDS1 immunity signaling. In a tpr1 tpl tpr4 (t3) mutant, resistance to bacteria was slightly compromised, and defense-related transcriptional reprogramming was weakly reduced or enhanced, respectively, at early (< 1 h) and late 24 h stages of bacterial infection. The t3 plants challenged with bacteria or pathogen-associated molecular pattern nlp24 displayed photosystem II dysfunctions. Also, t3 plants were hypersensitive to phytocytokine pep1 at the level of root growth inhibition. Transgenic expression of TPR1 rescued these t3 physiological defects. We propose that TPR1 and TPL family proteins function in Arabidopsis to reduce detrimental effects associated with activated transcriptional immunity.

Comparison of the Ways in Which Nitidine Chloride and Bufalin Induce Programmed Cell Death in Hematological Tumor Cells

The objective of this work to study the programmed cell death (PCD) in hematological tumor cells induced by nitidine chloride (NC) and bufalin (BF). Hematological tumor cells were exposed to various doses of NC and BF to measure the level of growth inhibition. While inverted microscope is used to observe cell morphology, western blot technique is used to detect apoptosis-related protein expression levels. The effects of NC and BF on hematological tumor cells were different. Although abnormal cell morphology could be seen under the inverted microscope, the western blot results showed that the two medicines induced PCD through different pathways. Drug resistance varied in intensity across distinct cells. THP-1, Jurkat, and RPMI-8226 each had half maximum inhibitory concentrations (IC50) of 36.23 nM, 26.71 nM, and 40.46 nM in BF, and 9.24 µM, 4.33 µM, and 28.18 µM in NC, respectively. Different hematopoietic malignancy cells exhibit varying degrees of drug resistance, and the mechanisms by which apoptosis of hematologic tumor cells is triggered by NC and BF are also distinct.

Insecticidal activity of Bacillus thuringiensis towards Agrotis exclamationis larvae-A widespread and underestimated pest of the Palearctic zone.

acillus thuringiensis is an entomopathogenic bacterium commonly used as a bioinsecticide against numerous invertebrate pests. However, the efficacy of this microbe has not yet been determined towards Agrotis exclamationis-a lepidopteran, polyphagous pest, widespread throughout the Palearctic zone. In this work we have detected very low susceptibility of A. exclamationis to B. thuringiensis commercial strains, used as microbial formulations in pest control. To investigate this matter, the biological activity of six selected (Cry1Aa, Cry1Ca, Cry1Ia, Cry2Ab, Cry9Ea and Vip3Aa), heterogously-expressed Bacillus thuringiensis insecticidal proteins has been assessed towards A. exclamationis. Only Cry9Ea and Vip3Aa caused significant mortality in the tested pest species, with LC50 values of 950 and 140 ng/cm2, respectively. The histopathological effects of Cry9Ea and Vip3Aa on A. exclamationis were determined. On the other hand, Cry1- and Cry2-type toxins, which are the main active molecules of the majority of currently-used B. thuringiensis-based biocontrol agents (including the commercial strains tested in this work), did not cause mortality in target insect, but only different levels of growth inhibition. Moreover, in the case of Cry1Ca and Cry1Ia hormesis has been observed-a phenomenon that may be disadvantageous in implementation of these proteins in pest management. The obtained results broaden the existing knowledge regarding B. thuringiensis insecticidal protein target range and depict variable susceptibility of A. exclamationis to different groups of Cry/Vip toxins. This work indicates Cry9Ea and Vip3Aa as good candidates for efficient biological control of A. exclamationis and possibly other Agrotinae and discusses the potential use of Vip3-type and Cry9-type insecticidal proteins as successful bioinsecticides.

Volatile Organic Compounds from Pythium oligandrum Play a Role in Its Parasitism on Plant-Pathogenic Pythium myriotylum.

The oomycete Pythium oligandrum is a soil-inhabiting parasite and predator of both fungi and oomycetes, and uses hydrolytic enzymes extensively to penetrate and hydrolyze its host or prey. Other mechanisms have been studied less, and we investigated the contribution of P. oligandrum-produced volatile organic compounds (VOCs) to parasitism. The growth-inhibiting activity of P. oligandrum VOCs was tested on Pythium myriotylum-a host or prey of P. oligandrum-coupled with electron microscopy, and biochemical and transcriptomic analyses. The P. oligandrum-produced VOCs reduced P. myriotylum growth by 80% and zoospore levels by 60%. Gas chromatography-mass spectrometry (GC-MS) identified 23 VOCs, and methyl heptenone, d-limonene, 2-undecanone, and 1-octanal were potent inhibitors of P. myriotylum growth and led to increased production of reactive oxygen species at a concentration that did not inhibit P. oligandrum growth. Exposure to the P. oligandrum VOCs led to shrinkage of P. myriotylum hyphae and lysis of the cellular membranes and organelles. Transcriptomics of P. myriotylum exposed to the P. oligandrum VOCs at increasing levels of growth inhibition initially showed a strong upregulation of putative detoxification-related genes that was not maintained later. The inhibition of P. myriotylum growth continued immediately after the exposure to the VOCs was discontinued and led to the reduced infection of its plant hosts. The VOCs produced by P. oligandrum could be another factor alongside hydrolytic enzymes contributing to its ecological role as a microbial parasite in particular ecological niches such as in soil, and may also contribute to the biocontrol of diseases using P. oligandrum commercial preparations. IMPORTANCE Microbe-microbe interactions in nature are multifaceted, with multiple mechanisms of action, and are crucial to how plants interact with microbes. Volatile organic compounds (VOCs) have diverse functions, including contributing to parasitism in ecological interactions and potential applications in biocontrol. The microbial parasite P. oligandrum is well known for using hydrolytic enzymes as part of its parasitism. We found that P. oligandrum VOCs reduced the growth of, and caused major damage to, the hyphae of P. myriotylum (a host or prey of P. oligandrum). Transcriptomic analyses of P. myriotylum exposed to the VOCs revealed the upregulation of genes potentially involved in an attempt to detoxify the VOCs. The inhibitory effects of the VOCs had a knock-on effect by reducing the virulence of P. myriotylum toward its plant hosts. The P. oligandrum VOCs could contribute to its ecological role as a microbial parasite. The VOCs analyzed here may also contribute to the biocontrol of diseases using P. oligandrum commercial preparations.

Chemical composition and anti-Helicobacter pylori activity of essential oil from fresh fruits of Litsea cubeba (Lour.) Pers

ABSTRACT Main chemical compositions of essential oil (EO) from Litsea cubeba fruit collected in Kon Tum Province of Vietnam were geranial (32.0%), neral (24.3%), citronellal (14.2%), and limonene (10.1%). Twenty-four compounds of the EO were identified and compared with those from EOs previously reported in India, China, and Malaysia. Minimal inhibitory and bactericidal concentrations (MIC and MBC) indicated that the EO induced high levels of growth inhibition and moderate bactericidal activity against seven test Helicobacter pylori strains, including six strains resistant to one to four antibiotics, azithromycin, erythromycin, levofloxacin, and metronidazole. The bacterial biofilm formation was significantly suppressed by the EO at 1/2 and 3/4×MIC. The EO also exerts inhibitory activity against H. pylori-urease and convert a spiral form of H. pylori into a coccoid form. Fruit EO inhibits bacterial growth via both contact and fumigant action.

Novel disulfide bond bridged 7-ethyl-10-hydroxyl camptothecin-undecanoic acid conjugate/human serum albumin nanoparticles for breast cancer therapy.

7-Ethyl-10-hydroxyl camptothecin (SN38), a semisynthetic derivative of camptothecin, exhibited extreme pharmacological activities in treating a range of cancers. However, its poor aqueous solubility and low stability hinder its clinical applications. Hence, a redox-responsive SN38 prodrug encapsulated human serum albumin (HSA) nanoparticle is developed to realize its potential in the clinic. First, a disulfide bond bridged 7-ethyl-10-hydroxyl camptothecin-undecanoic acid conjugate (SN38-SS-COOH) was synthesized and characterized structurally. After that, SN38-SS-COOH/HSA nanoparticles (SNH NPs) were prepared by the desolvation method. The SNH NPs with a feed molar ratio of 9 : 1 of SN38-SS-COOH : HSA showed a spherical structure with a diameter range of approximately 120-150 nm revealed by dynamic light scattering (DLS) and transmission electron microscopy (TEM). Fluorescence quenching confirmed the formation of SNH NP complexes by dual hydrophobic force and electrostatic interaction. The SNH NPs have a high drug loading of 10.44% and an encapsulation efficiency of 89.59% with good stability. Moreover, the redox responsiveness was validated by glutathione (GSH)-triggered accelerated release of parent drug SN38. In an in vivo pharmacokinetic study, the SNH NPs exhibited a significantly prolonged circulation time (t1/2, 3.77-fold) compared with free SN38. Finally, the in vivo antitumor efficacy and systemic toxicity of SNH NPs in a breast xenograft model were thoroughly evaluated. The inhibition rate of tumor growth induced by the SNH NPs reached 70.1%, while only 50.1% was achieved for irinotecan at an equivalent SN38 dosage of 10 mg kg-1. More importantly, the SNH NPs achieved a higher level of tumor growth inhibition (85.3%) by increasing the dosage to 60 mg kg-1 SN38 without obvious adverse effects. Taken together, the use of redox-responsive SN38 prodrug/HSA NPs could be a promising strategy to deliver highly active SN38 for breast cancer chemotherapy.

Synthesis and Preclinical Evaluation of Small-Molecule Prostate-Specific Membrane Antigen-Targeted Abiraterone Conjugate.

Prostate cancer is the second most common type of cancer among men. The main method of its treatment is androgen deprivation therapy, which has a wide range of side effects. One of the solutions to this challenge is the targeted delivery of drugs to prostate cancer cells. In this study, we performed the synthesis of a novel small-molecule PSMA-targeted conjugate based on abiraterone. Cytotoxicity, the induction of intracellular reactive oxygen species, and P450-cytochrome species inhibition were investigated for this conjugate PSMA-abiraterone. The conjugate demonstrated a preferential effect on prostate tumor cells, remaining inactive at up to 100 µM in human fibroblast cells. In addition, it revealed preferential efficacy, specifically on PSMA-expressing lines with a 65% tumor growth inhibition level on 22Rv1 (PSMA+) xenografts after 14-fold oral administration of PSMA-Abi at a single dose of 500 mg/kg (7.0 g/kg total dose) was observed. This compound showed significantly reduced acute toxicity with comparable efficacy compared to AbiAc.